title: Effects of culture and transplantation on follicle activation and early follicular growth in neonatal mouse ovaries.
abstract: Mouse models have been widely utilized to elucidate the basic principles and regulatory mechanisms of primordial follicle activation. Outside their natural environment, the growth of follicles might be affected by unknown factors in vitro and the elimination of regulation in vivo. Currently, in vitro culture and transplantation of ovaries under the kidney capsule are two commonly used incubation methods. However, the limited number of studies that have been published compare various incubation systems and reveal differences between ovaries that are incubated and grown in vivo. We compare the number of primordial, primary and secondary follicles in cultured, transplanted and in-vivo-grown ovaries. We investigate the expression levels of four genes, including zona pellucida 3 (ZP3), growth and differentiation factor-9 (GDF-9), proliferating cell nuclear antigen (PCNA) and anti-Müllerian hormone (AMH). Our results suggest that in vitro culture accelerates follicle activation, delays the transition from primary to secondary follicles and affects the expression patterns of ZP3, GDF-9, PCNA and AMH. A larger number of secondary follicles in ovaries cultured in alpha-minimal essential medium (α-MEM) had intact zona pellucida compared with those grown in Dulbecco's modified Eagle medium containing Ham's F-12 nutrient mixture (D/F12), suggesting that α-MEM is a better basal medium. The transplanted ovaries demonstrated the most similar characteristics to the in-vivo-grown ovaries, indicating that transplantation provided an optimal environment for ovarian incubation. This study has thus established the similarities and differences between in-vivo-grown and incubated ovaries, demonstrated that transplantation can mostly mimic the environment of ovarian growth in vivo and determined the optimal basal culture medium between α-MEM and D/F12.
authors.affiliation: Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, 430030, People's Republic of China.
authors.forename: Shuo
authors.initials: S
authors.lastname: Wang
authors.forename: Shuhong
authors.initials: S
authors.lastname: Yang
authors.forename: Zhiwen
authors.initials: Z
authors.lastname: Lai
authors.forename: Ting
authors.initials: T
authors.lastname: Ding
authors.forename: Wei
authors.initials: W
authors.lastname: Shen
authors.forename: Liangyan
authors.initials: L
authors.lastname: Shi
authors.forename: Jingjing
authors.initials: J
authors.lastname: Jiang
authors.forename: Lanfang
authors.initials: L
authors.lastname: Ma
authors.forename: Yong
authors.initials: Y
authors.lastname: Tian
authors.forename: Xiaofang
authors.initials: X
authors.lastname: Du
authors.forename: Aiyue
authors.initials: A
authors.lastname: Luo
authors.forename: Shixuan
authors.initials: S
authors.lastname: Wang
mesh_terms: D000818:Animals; D005260:Female; D018507:Gene Expression Regulation, Developmental; D006107:Granulosa Cells / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D051379:Mice; D008810:Mice, Inbred C57BL; D009865:Oocytes / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D009924:Organ Culture Techniques; D006080:Ovarian Follicle / Q000254:growth & development / Q000378:metabolism / Q000648:ultrastructure; D010053:Ovary / Q000254:growth & development* / Q000378:metabolism / Q000637:transplantation* / Q000648:ultrastructure